Miroestrol (ME) is a potent phytoestrogen from the P. candollei tuberous
root. It has been approved for use in clinical trials due to its
beneficial effect on disorders associated with estrogen deficiency. To
ensure medical efficacy and safety, high performance analytical methods
for ME analysis are required to standardize products from the P.
candollei root. An enhanced chemiluminescence enzyme-linked
immunosorbent assay (ECL-ELISA) was developed and validated using a
polyclonal antibody against ME and a chemiluminescent system of
luminol-H2 O2 -horseradish peroxidase-4-(1-imidazolyl) phenol. The
ECL-ELISA system exhibited linearity over a concentration range of
0.31-10.00 ng mL(-1) , for which the relative standard variation (%RSD)
was less than 10% for both intra- and interplate determinations. The
ECL-ELISA is reliable for the determination of ME as reflected by the
high recovery percentage (101.22-103.06%). As a comparative analysis,
the ME content in each sample determined by ECL-ELISA was correlated
with high coefficients of determination with colorimetric ELISA (R(2)
= 0.998) and high performance liquid chromatography (HPLC) (R(2)
= 0.998) methods. The ECL-ELISA method could be applied to all of the
commercial products containing P. candollei root, when the products
contain between 0.706 ± 0.046 and 13.123 ± 0.794 µg g(-1) dry wt. of ME.
This method is useful as a high performance analytical method for the
quantity control of ME in raw materials and end products at both the
research and industrial levels.